This invention relates to 4-(4-substituted-benzenesulfonyl)-2,3,4,5-tetrahydro-1H-[1,4]benzodiazepine-3-hydroxamic acids which act as matrix metalloproteinase inhibitors and as inhibitors of TNF-xcex1 converting enzyme(TACE). The compounds of the present invention are useful in disease conditions mediated by matrix metalloproteinases and TACE, such as tumor growth, osteoarthritis, rheumatoid arthritis and degenerative cartilage loss.
Matrix metalloproteinases (MMPs) are a group of enzymes that have been implicated in the pathological destruction of connective tissue and basement membranes. These zinc-containing endopeptidases consist of several subsets of enzymes, including collagenases, stromelysins and gelatinases. Of these, the gelatinases have been shown to be the MMPs most intimately involved with the growth and spread of tumors.
For example, it is known that the level of expression of gelatinase is elevated in malignancies, and that gelatinase can degrade the basement membrane which leads to tumor metastasis. Angiogenesis, required for the growth of solid tumors, has also recently been shown to have a gelatinase component to its pathology as reported in xe2x80x9cMatrix Metalloproteinases, Novel Targets for Directed Cancer Therapyxe2x80x9d, Drugs and Aging, 11:229-244 (1997).
Other conditions mediated by MMPs include restenosis, MMP-mediated osteopenias, inflammatory diseases of the central nervous system, skin aging, osteoarthritis, rheumatoid arthritis, septic arthritis, corneal ulceration, abnormal wound healing, bone disease, proteinuria, aneurysmal aortic disease, degenerative cartilage loss following traumatic joint injury, demyelinating diseases of the nervous system, cirrhosis of the liver, glomerular disease of the kidney, premature rupture of fetal membranes, inflammatory bowel disease, periodontal disease, age-related macular degeneration, diabetic retinopathy, proliferative vitreoretinopathy, retinopathy of prematurity, ocular inflammation, keratoconus, Sjogren""s syndrome, myopia, ocular tumors, ocular angiogenesis/ neo-vascularization and corneal graft rejection. Studies relating to these conditions are set forth, e.g., in xe2x80x9cRecent Advances in Matrix Metalloproteinase Inhibitor Researchxe2x80x9d, R. P. Beckett et al., Research Focus, 1:16-26, (1996); Curr. Opin. Ther. Patents. 4(1): 7-16, (1994); Curr. Medicinal Chem., 2: 743-762, (1995); Exp. Opin. Ther. Patents, 5(2): 1087-110, (1995); Exp. Opin. Ther. Patents, 5(12): 1287-1196, (1995); xe2x80x9cInhibition of Matrix Metallo-proteinases: Structure Based Designxe2x80x9d, Current Pharmaceutical Design, 2:524-661, (1996). xe2x80x9cMatrix Metalloproteinase Inhibitor Drugsxe2x80x9d, Emerging Drugs, 2:205-230 (1997).
TNF-xcex1 converting enzyme (TACE) catalyzes the formation of TNF-xcex1 from membrane bound TNF-xcex1 precursor protein. TNF-xcex1 is a pro-inflammatory cytokine that is believed to have a role in rheumatoid arthritis, septic shock, graft rejection, cachexia, anorexia, inflammation, congestive heart failure, inflammatory disease of the central nervous system, inflammatory bowel disease, insulin resistance and HIV infection, in addition to its well-documented antitumor properties. Research with anti-TNF-xcex1 antibodies in transgenic animals has demonstrated that blocking the formation of TNF-xcex1 inhibits the progression of arthritis. This observation has recently been extended to humans as described in xe2x80x9cTNF-xcex1 in Human Diseasesxe2x80x9d, Current Pharmaceutical Design, 2:662-667 (1996).
It is expected that small molecule inhibitors of MMPs and TACE would have the potential for treating a variety of disease states. Although a variety of MMP and TACE inhibitors are known, many of these molecules are peptidic and peptide-like which demonstrate bioavailability and pharmacokinetic problems. Long acting, orally bioavailable non-peptide inhibitors of MMPs and/or TACE would thus be highly desirable for the treatment of the disease states discussed above.
U.S. Pat. No. 5,455,258 discloses 2-substituted-2-(arylsulfonylamino) hydroxyamic acids and their use as MMP inhibitors. WO 97/18194, discloses N-(arylsulfonyl)tetrahydroisoquinolone-hydroxamic acids and related bicyclic derivatives thereof and their use as MMP inhibitors. WO 97/20824 and U.S. Pat. No. 5,753,653 disclose 1-(arylsulfonyl)-4-(substituted)piperazine-2-hydroxamic acids,
4-(arylsulfonyl)morpholine-3-hydroxamic acids, 4-(arylsulfonyl)-tetrahydro-2H,1,4-thiazine-3-hydroxamic acids, 3-(substituted-1-(arylsulfonyl)hexahydro-2-hydroxamic acids and related compounds as useful MMP inhibitors.
WO 98/08822, WO 98/08823 and WO 98/08825, disclose 6-membered 1-(arylsulfonyl)hexahydropyrimidine-2-hydroxamic acids, 1-substituted-3-[(4-methoxybenzenesulfonyl)]hexahydropyrimidine-4-hydroxamic acids, 4-(arylsulfonyl)-tetrahydro-1,2-thiazine-3-hydroxamic acids and (arylsulfonyl)-4-substitutedpiperazine-2-hydroxamic acids. WO 98/08827 discloses 4-(arylsulfonyl)-hexahydrothiazepine-3-hydroxamic acids and 4-(arylsulfonyl)-hexadydro[1,4]diazepine-3-hydroxamic acids.
This invention relates to novel derivatives of substituted 2,3,4,5-tetrahydro-1H-[1,4]benzodiazepine-3-carboxylic acid, hydroxamide which exhibit inhibitory activity against MMPs. The compounds of the present invention are represented by the following formula 1

 
wherein
R is selected from hydrogen, (C1-C3)alkyl, xe2x80x94CN, xe2x80x94ORxe2x80x2, xe2x80x94SRxe2x80x2, xe2x80x94CF3, xe2x80x94OCF3, Cl, F, NH2, NH(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)CO(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)(Rxe2x80x2), NO2, xe2x80x94CONH2, xe2x80x94SO2NH2, xe2x80x94SO2N(Rxe2x80x2)(Rxe2x80x2), or xe2x80x94N(Rxe2x80x2)COCH2Oxe2x80x94(C1-C3)alkyl, wherein Rxe2x80x2 is (C1-C3) alkyl or hydrogen;
R4 is (C2-C6) alkyl-Oxe2x80x94 containing one triple bond;

 
wherein Rxe2x80x3 is hydrogen, xe2x80x94CH2OH, (C1-C6)alkyl, (C1-C6)alkyl-Oxe2x80x94CH2xe2x80x94, (C1-C6)alkyl-Sxe2x80x94CH2xe2x80x94, (C1-C6)alkyl-NHxe2x80x94CH2xe2x80x94, [(C1-C3)alkyl]2-NCH2xe2x80x94, (C1-C6)oydodkyl-Oxe2x80x94CH2xe2x80x94, ((C1-C3)dkyl)2-Nxe2x80x94(CH22-4NHCH2xe2x80x94, [(C1-C3)dkyl)2-Nxe2x80x94(CH2)2-4N(CH3)CH2xe2x80x94,

 

 
R1 and R2 are each, independently, hydrogen or C1-C3alkyl; R3 is (C1-C8)alkyl, NH2CH2COxe2x80x94, (C1-C6)alkylNHCH2COxe2x80x94, HO(CH2)mCOxe2x80x94, HCOxe2x80x94, Aryl(CH2)nCOxe2x80x94, Heteroaryl(CH2)nCOxe2x80x94, (C1-C3)alkyl-Oxe2x80x94(CH2)nCOxe2x80x94, (C1-C3)alkylCOxe2x80x94, (C1-C3)alkylCOxe2x80x94NHCH2COxe2x80x94, (C3-C7)cycloalkylCOxe2x80x94, (C1-C3)alkylSO2xe2x80x94, Aryl(CH2)nSO2xe2x80x94, Heteroaryl(CH2)nSO2xe2x80x94, (C1-C3)alkyl-Oxe2x80x94(CH2)mxe2x80x94SO2xe2x80x94, (C1-C3)alkyl-Oxe2x80x94(CH2)mxe2x80x94, (C1-C3)alkyl-Oxe2x80x94(C1-C3)alkyl-Oxe2x80x94(C1-C3)alkyl, HOxe2x80x94(C1-C3)alkyl-Oxe2x80x94(C1-C3)alkyl, Aryl-Oxe2x80x94CH2COxe2x80x94, Heteroaryl-Oxe2x80x94CH2COxe2x80x94, ArylCHxe2x95x90CHCOxe2x80x94, HeteroarylCHxe2x95x90CHCOxe2x80x94, (C1-C3)alkylCHxe2x95x90CHCOxe2x80x94,

 
Aryl(C1-C3)alkyl, Heteroaryl(C1-C3)alkyl, ArylCHxe2x95x90CHCH2xe2x80x94,
HeteroarylCHxe2x95x90CHCH2xe2x80x94, (C1-C6)alkylCHxe2x95x90CHCH2xe2x80x94,

 
Rxe2x80x2OCH2CH(ORxe2x80x2)COxe2x80x94, (Rxe2x80x2OCH2)2C(Rxe2x80x2)COxe2x80x94,

 
[(C1-C6)alkyl]2-Nxe2x80x94(C1-C6)alkyl COxe2x80x94, or (C1-C6)alkyl-NHxe2x80x94(C1-C6)alkylCOxe2x80x94;
wherein
m=1 to 3; n=0 to 3;
Aryl is

 
Heteroaryl is

 
wherein X is hydrogen, halogen, (C1-C3) alkyl or xe2x80x94OCH3 and R and Rxe2x80x2 are as defined above;
L is hydrogen, (C1-C3)alkyl, xe2x80x94CN, xe2x80x94ORxe2x80x2, xe2x80x94SRxe2x80x2, xe2x80x94CF3, xe2x80x94OCF3, Cl, F, NH2, xe2x80x94NHxe2x80x94(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)CO(C1-C3)alkyl, N(Rxe2x80x2)(Rxe2x80x2), xe2x80x94NO2, xe2x80x94CONH2, xe2x80x94SO2NH2, xe2x80x94SO2N(Rxe2x80x2)(Rxe2x80x2), xe2x80x94N(Rxe2x80x2)COCH2Oxe2x80x94(C1-C3)alkyl,

 
M is

 

 
xe2x80x83or N(Rxe2x80x2)(Rxe2x80x2) where Rxe2x80x2 is as defined above;
W is O, S, NH or N(C1-C3)alkyl;
Y is hydrogen, F, Cl, CF3 or OCH3; and Xxe2x80x2 is halogen, hydrogen, (C1-C3)alkyl, Oxe2x80x94(C1-C3)alkyl, or xe2x80x94CH2OH; and
pharmaceutically acceptable salts thereof.
Preferably, the compounds of the present invention are those of formula 1 wherein R is hydrogen, (C1-C3) alkyl, xe2x80x94CN, xe2x80x94ORxe2x80x2, xe2x80x94SRxe2x80x2, xe2x80x94CF3, xe2x80x94OCF3, Cl, F, NH2, NH(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)CO(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)(Rxe2x80x2), NO2, xe2x80x94CONH2, xe2x80x94SO2NH2, xe2x80x94SO2N(Rxe2x80x2)(Rxe2x80x2), or xe2x80x94N(Rxe2x80x2)COCH2Oxe2x80x94(C1-C3)alkyl, wherein Rxe2x80x2 is (C1-C3) alkyl or hydrogen;
R4 is (C1-C6) alkyl-Oxe2x80x94 containing one triple bond,

 

 
xe2x80x83xe2x80x94Oxe2x80x94CH2xe2x80x94Cxe2x89xa1Cxe2x80x94Rxe2x80x3
wherein Rxe2x80x3 is hydrogen , xe2x80x94CH2OH, (C1-C6)alkyl, (C1-C6)alkyl-Oxe2x80x94CH2xe2x80x94, (C1-C6)akyl-Sxe2x80x94CH2xe2x80x94, (C1-C6)alkyl-NHxe2x80x94CH2xe2x80x94, [(C1-C3)alkyl]2-NCH2xe2x80x94, (C1-C6)cycloalkyl-Oxe2x80x94CH2xe2x80x94, [(C1-C3)alkyl]2-Nxe2x80x94(CH2)2-4NHCH2xe2x80x94, [(C1-C3)alkyl]2-Nxe2x80x94(CH2)2-4N(CH3)CH2xe2x80x94,

 

 
R1 and R2 are each, independently, hydrogen or CH3;
R3 is (C1-C8)alkyl, NH2CH2COxe2x80x94, (C1-C6)alkylNHCH2COxe2x80x94, HO(CH2)mCOxe2x80x94, HCOxe2x80x94, Aryl(CH2)nCOxe2x80x94, Heteroaryl(CH2)nCOxe2x80x94, (C1-C3)alkyl-Oxe2x80x94(CH2)nCOxe2x80x94, (C1-C3)alkylCOxe2x80x94, (C1-C3)alkylCOxe2x80x94NHCH2COxe2x80x94, (C3-C7)cycloalkylCOxe2x80x94, Aryl-Oxe2x80x94CH2COxe2x80x94, HeteroarylOCH2COxe2x80x94,

 
wherein
m=1 to 3; n=0 to 3;
Aryl is

 
Heteroaryl is

 
wherein X is hydrogen, halogen, (C1-C3)alkyl, or xe2x80x94OCH3 wherein R and Rxe2x80x2 are as defined above; and pharmaceutically acceptable salts thereof.
It is more preferred that the compounds of the present invention include those of formula 1 wherein R is hydrogen, (C1-C3) alkyl, xe2x80x94CN, xe2x80x94ORxe2x80x2, xe2x80x94SRxe2x80x2, xe2x80x94CF3, xe2x80x94OCF3, Cl, F, NH2, NH(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)CO(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)(Rxe2x80x2), NO2, xe2x80x94CONH2, xe2x80x94SO2NH2, xe2x80x94SO2N(Rxe2x80x2)(Rxe2x80x2), or xe2x80x94N(Rxe2x80x2)COCH2Oxe2x80x94(C1-C3)alkyl, wherein Rxe2x80x2 is (C1-C3) alkyl or hydrogen;
R4 is (C1-C6) alkyl-Oxe2x80x94 containing one triple bond,

 
wherein Rxe2x80x3 is hydrogen , xe2x80x94CH2OH (C1-C6)alkyl, (C1-C6)alkyl-Oxe2x80x94CH2xe2x80x94, (C1-C6)alkyl-Sxe2x80x94CH2xe2x80x94, (C1-C6)alkyl-NHxe2x80x94CH2xe2x80x94, [(C1-C3)alkyl]2-NCH2xe2x80x94, (C1-C6)cycloalkyl-Oxe2x80x94CH2xe2x80x94, [(C1-C3) alkyl]2-Nxe2x80x94(CH2)2-4NHCH2xe2x80x94, [(C1-C3) alkyl]2-Nxe2x80x94(CH2)2-4N(CH3)CH2xe2x80x94,

 

 
R1 and R2 are each, independently hydrogen or CH3;
R3 is (C1-C3)alkylCOxe2x80x94, (C1-C3)alkyl-Oxe2x80x94(CH2)mCOxe2x80x94, ArylCOxe2x80x94
wherein
m=1 to 3; n=0 to 3;

 
Aryl is
Heteroaryl is

 
wherein X is hydrogen, halogen, (C1-C3) alkyl or xe2x80x94OCH3 and R and Rxe2x80x2 are as defined above; and pharmaceutically acceptable salts thereof.
It is more preferred that the compounds of the present invention include those of formula 1 wherein R is hydrogen, (C1-C3) alkyl,xe2x80x94CN,xe2x80x94OR,xe2x80x94SR,xe2x80x94CF3,OCF3,Cl, F, NH2, NH (C1-C3) alkyl, xe2x80x94N(Rxe2x80x2)CO(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)(Rxe2x80x2), NO2,xe2x80x94CONH2,xe2x80x94SO2NH2, xe2x80x94SO2 NH2, SO2N(Rxe2x80x2)(Rxe2x80x2) or xe2x80x94N(Rxe2x80x2) COCHOxe2x80x94(C1-C3) alkyl, wherein Rxe2x80x2 is (C1-C3)alkyl or hydrogen;
R4 is (C1-C6) alkyl-Oxe2x80x94 containing one triple bond

 
wherein Rxe2x80x3 is hydrogen , CH2OH ,(C1-C6)alkyl, (C1-C6)alkyl-Oxe2x80x94CH2xe2x80x94, (C1-C6)alkyl-Sxe2x80x94CH2xe2x80x94, (C1-C6)alkyl-NHxe2x80x94CH2xe2x80x94, [(C1-C3)alkyl]2-NCH2xe2x80x94, (C1-C6)cycloalkyl-Oxe2x80x94CH2xe2x80x94, [(C1-C3) alkyl]2-Nxe2x80x94(CH2)2-4NHCH2xe2x80x94, [(C1-C3) alkyl]2-Nxe2x80x94(CH2)2-4N(CH3)CH2xe2x80x94,

 

 
R1 and R2 are each, independently hydrogen or CH3 
R3 is (C1-C3) alkylSO2-Aryl (CH2)nSO2xe2x80x94, Heteroary(CH2)nSO2xe2x80x94, or (C1-C3) alkyl-Oxe2x80x94(CH2)nSO2xe2x80x94,
wherein
m=1 to 3; n=0 to 3;
Aryl is

 
Heteroaryl is

 
wherein X is hydrogen, halogen, (C1-C3) alkyl or xe2x80x94OCH3 and R and Rxe2x80x2 are as defined above; and pharmaceutically acceptable salts thereof.
It is more preferred that the compounds of the present invention include those of formula 1 wherein R is hydrogen, (C1-C3) alkyl,xe2x80x94CN,xe2x80x94OR,xe2x80x94SR,xe2x80x94CF3,OCF3,,Cl, F, NH2, NH (C1-C3) alkyl, xe2x80x94N(Rxe2x80x2)CO(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)(Rxe2x80x2), NO2,xe2x80x94CONH2,xe2x80x94SO2NH2, xe2x80x94SO2NH2, SO2N(Rxe2x80x2)(Rxe2x80x2) or xe2x80x94N(Rxe2x80x2) COCH2Oxe2x80x94(C1-C3) alkyl, wherein Rxe2x80x2 is (C1-C3)alkyl or hydrogen;
R4 is (C1-C6) alkyl-Oxe2x80x94 containing one triple bond
A further, more preferred embodiment of the present invention includes compounds represented by formula 1 wherein
R is selected from hydrogen, (C1-C3) alkyl, xe2x80x94CN, xe2x80x94ORxe2x80x2, xe2x80x94SRxe2x80x2, xe2x80x94CF3, xe2x80x94OCF3, Cl, F, NH2, NH(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)CO(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)(Rxe2x80x2), NO2, xe2x80x94CONH2, xe2x80x94SO2NH2, xe2x80x94SO2N(Rxe2x80x2)(Rxe2x80x2), or xe2x80x94N(Rxe2x80x2)COCH2Oxe2x80x94(C1-C3)alkyl, wherein Rxe2x80x2 is (C1-C3) alkyl or hydrogen;
R4 is (C1-C6)alkyl-Oxe2x80x94 containing one triple bond,

 
wherein Rxe2x80x3 is hydrogen , xe2x80x94CH2OH, (C1-C6)alkyl, (C1-C6)alkyl-Oxe2x80x94CH2xe2x80x94, (C1-C6)alkyl-Sxe2x80x94CH2xe2x80x94, (C1-C6)alkyl-NHxe2x80x94CH2xe2x80x94, [(C1-C3)alkyl]2-NCH2xe2x80x94, (C1-C6)cycloalkyl-Oxe2x80x94CH2xe2x80x94, [(C1-C3)alkyl]2-Nxe2x80x94(CH2)2-4NHCH2xe2x80x94, [(C1-C3)alkyl]2-Nxe2x80x94(CH2)2-4N(CH3)CH2xe2x80x94,

 

 
R1 and R2 are each, independently hydrogen or CH3; R3 is (C1-C3) alkylCOxe2x80x94, (C1-C3)alkyl-COxe2x80x94, (C1-C7)cycloalkyCOxe2x80x94, (C1-C3)alkyl-Oxe2x80x94(CH2)mxe2x80x94COxe2x80x94, Ar (CH2)nCOxe2x80x94, HOxe2x80x94(CH2)mCOxe2x80x94, Heteroaryl(CH2)mxe2x80x94COxe2x80x94
wherein
Aryl is

 
Heteroaryl is

 
wherein X is hydrogen, halogen, (C1-C3) alkyl or xe2x80x94OCH3 and R and Rxe2x80x2 are as defined above; and pharmaceutically acceptable salts thereof.
Additionally highly preferred compounds of the present invention include those of formula 1 wherein R is hydrogen, (C1-C3) alkyl, xe2x80x94CN, xe2x80x94ORxe2x80x2, xe2x80x94SRxe2x80x2, xe2x80x94CF3xe2x80x94OCF3, Cl, F, NH2, NH(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)CO(C1-C3)alkyl, xe2x80x94N(Rxe2x80x2)(Rxe2x80x2), NO2, xe2x80x94CONH2, xe2x80x94SO2NH2, xe2x80x94SO2N(Rxe2x80x2)(Rxe2x80x2), or xe2x80x94N(Rxe2x80x2)COCH2Oxe2x80x94(C1-C3)alkyl, wherein Rxe2x80x2 is (C1-C3) alkyl or hydrogen;
R4 is xe2x80x94Oxe2x80x94CH2xe2x80x94Cxe2x89xa1Cxe2x80x94Rxe2x80x3;
wherein Rxe2x80x3 is hydrogen , xe2x80x94CH2OH , (C1-C6)alkyl , (C1-C6)alkyl-Oxe2x80x94CH2xe2x80x94, (C1-C6)alkyl-Sxe2x80x94CH2xe2x80x94, (C1-C6)alkyl-NHxe2x80x94CH2xe2x80x94, [(C1-C3)alkyl]2-NCH2xe2x80x94, (C1-C6)cycloalkyl-Oxe2x80x94CH2xe2x80x94, [(C1-C3) alkyl]2-Nxe2x80x94(CH2)2-4NHCH2xe2x80x94, [(C1-C3) alkyl]2-Nxe2x80x94(CH2)2-4N(CH3)CH2xe2x80x94,

 

 
R1 and R2 are each, independently hydrogen or CH3;
R3 is

 
wherein
m=1 to 3; n=0 to 3;
L is hydrogen, (C1-C3)alkyl, xe2x80x94CN, xe2x80x94ORxe2x80x2, xe2x80x94SRxe2x80x2, xe2x80x94CF3, xe2x80x94OCF3, Cl, F, NH2, xe2x80x94NHxe2x80x94(C1-C3)alkyl, N(Rxe2x80x2)CO(C1-C3)alkyl, N(Rxe2x80x2)(Rxe2x80x2), xe2x80x94NO2, xe2x80x94CONH2, xe2x80x94SO2NH2, xe2x80x94SO2N(Rxe2x80x2)(Rxe2x80x2), xe2x80x94N(Rxe2x80x2)COCH2Oxe2x80x94(C1-C3)alkyl,

 
M is

 

 
xe2x80x83or N(Rxe2x80x2)(Rxe2x80x2) where Rxe2x80x2 is as defined above;
W is O, S, NH or N(C1-C3)alkyl;
Y is hydrogen, F, Cl, CF3 or OCH3; and Xxe2x80x2 is halogen, hydrogen, (C1-C3)alkyl, Oxe2x80x94(C1-C3)alkyl, or xe2x80x94CH2OH; and pharmaceutically acceptable salts thereof.
Some of the intermediate compounds for the preparation of derivatives of formula 1 are those wherein R4 is OCH3 and they may be advantageously prepared according to the illustrated Reaction Schemes. Ester derivatives of formulae 8,14, 15,20,22,23,24,25,29 and 30 wherein R4 is OCH3 are used to prepare derivatives wherein R4 is OH (via cleavage of Oxe2x80x94CH3 group). As illustrated in Scheme 8 derivatives 40 with a phenolic OH group are reacted under standard organic synthetic conditions to convert the OH moiety into the R4 substituents (as previously defined). Variations in these schemes may be made to improve productivity without negatively impacting the amount and nature of the product, by means that will be recognized by those skilled in the art. For example, reactive groups may be blocked with suitable blocking moieties which may then be deblocked under standard conditions (for instance, hydroxy groups may be protected with trimethylsilyl or t-butyl-dimethylsilyl moieties which are then removed in a later reaction step). In addition, those skilled in the art will recognize that catalylic hydrogenation conditions are inappropiate for preparing intermediates with an R4 moiety containing a triple bond; the R4 moiety is then introduced into intermediates not requiring a reduction step.
In general, the compounds of Formula 1 are synthesized from an alkyl ester (such as methyl, ethyl, t-butyl and the like) of serine, threonine, or 3,3-dimethyl-3-hydroxypropionic acids. One reaction pathway is shown in Reaction Scheme 1. It is noted that methyl esters are shown in all of the Reaction Schemes, however, it is to be understood that the use of methyl esters is for purposes of illustration only, and other suitable alkyl esters. benzyl esters and the like may similarly be used.
In Reaction Scheme 1, serine, threonine, beta-hydroxyvaline and related derivatives are converted to the corresponding N-(4-substituted-benzenesulfonyl) derivatives 3 and alkylated with suitable substituted or unsubstituted 2-nitrobenzyl bromides or 2-nitrobenzyl chlorides to provide the corresponding nitro derivatives 5 . Reduction under conventional reducing conditions, such as catalytic hydrogeneration (with Pd/C) or chemical reduction (e.g., with SnCl2 or FeCl3) results in amino derivatives 6. Reaction of the N-(2-aminobenzyl) derivatives 6 with alkanoyl chlorides, alkylsulfonyl chlorides, aroyl chlorides, heteroaroyl chlorides, aryl sulfonyl chlorides, heteroarylsulfonyl chlorides and the like, in the presence of trialkylamines or pyridene, provides the dehydroalanine derivatives 7. Ring closure to the [1,4]benzodiazepine compounds 9 is carried out by reaction with a mild base such as sodium or potassium bicarbonate in an alcohol solvent such as methanol or ethanol. Standard conditions which involve hydrolysis of the ester (NaOH), acid chloride formation and reaction of the acid chloride with hydroxylamine are then used to convert the ester derivatives 8 to the hydroxamic acids 9. Ester derivatives 8 (where the ester function is a t-butyl ester) are converted to the acid with trifluoroacetic acid under standard conditions.
As illustrated in Reaction Scheme 2, derivatives 10, which contain a blocked hydroxyl group, are alkylated with 2-nitro or 2-amino benzyl alcohol derivatives 11 by application of the Mitsunobu reaction to give intermediates 12. Reduction of the 2-nitro group and removal of the hydroxy blocking group with derivatives 12, where the R4 group is a protected amino moiety with simultaneous deblocking of the amino and hydroxyl functions, gives intermediate compounds 13. The intermediates 13 may then be reacted with benzyloxycarbonyl chloride to give the closed ring [1,4] benzodiazepines 14. Reaction of the derivatives 14 with acyl chlorides, aroyl chlorides, heteroaroyl chlorides, alkysulfonyl chlorides, arylsulfonyl chlorides and heteroarylsulfonyl chlorides and the like affords the intermediates 15.

 
wherein
n=0to 3;
m=1 to 3;
R1 and R2 are independently hydrogen or (C1-C3)alkyl; R=Hydrogen; halogen; OCH3; NO2; NH2; CF3; NHCOCH3; NHCOCH2CH3; CONH2; xe2x80x94N(Rxe2x80x2)(Rxe2x80x2), xe2x80x94N(Rxe2x80x2)CO(C1-C3)alkyl; (C1-C3)alkyl;
R3=Ar(CH2)nCOxe2x80x94; Heteroaryl(CH2)nCOxe2x80x94, Ar(CH2)nSO2xe2x80x94; Heteroaryl(CH2)nSO2xe2x80x94; Alkyl-Oxe2x80x94CH2)nCOxe2x80x94; Alkyl-Oxe2x80x94(CH2)mSO2xe2x80x94; AlkylCOxe2x80x94; AlkylSO2xe2x80x94; AlkylCOxe2x80x94NHCH2COxe2x80x94; and cycloalkyl(C3-C7)COxe2x80x94; and
R4 is as defined herein.

 
wherein
n=0 to 3;
m=1 to 3;
Ø=phenyl;
DEAD=diethylazodicarboxylate;
R6=Ar(CH2)nxe2x80x94; Alkyl-; Heteroaryl(CH2)nxe2x80x94; Alkyl-Oxe2x80x94(CH2)nxe2x80x94; Cycloalkyl(C3-C7);
R7=Ar(CH2)nxe2x80x94; Alkyl-; Heteroaryl(CH2)nxe2x80x94; Alkyl-Oxe2x80x94(CH2)mxe2x80x94;
R8=Ar(CH2)nCOxe2x80x94; Ar(CH2)nSO2xe2x80x94; AlkylCOxe2x80x94; AlkylSO2xe2x80x94; Heteroaryl(CH2)nCOxe2x80x94;
Heteroaryl(CH2)nSO2xe2x80x94; Alkyl-Oxe2x80x94(CH2)nCOxe2x80x94; Alkyl-Oxe2x80x94(CH2)mSO2xe2x80x94.
1-substituted arylmethyl-2,3,4,5-tetrahydro-1H [1,4]-benzodiazepines may be prepared in the manner illustrated in Reaction Schemes 3 and 4. In Reaction Scheme 3, the methyl 3-hydroxy-2-[4-methoxybenzenesulfonyl)-(2-amino-benzyl)amino]- propionates 6 are subjected to reductive alkylation with arylcarboxaldehydes and heteroarylcarboxaldehydes to provide intermediates 17. Standard reaction conditions such as reactions with triphenylphosphine and diethyl azodicarboxylate (DEAD) or triplenylphosphine with either carbon tetrachloride or carbon tetrabromide, results in the xe2x80x9cdehydroalaninexe2x80x9d derivatives 18 which are then ring closed to the [1,4]benzodiazepines 20.
In an alternative route to the 3-hydroxamic acid derivatives 21 (Scheme 4), N-aroyl derivatives 22 are reduced with reducing agents such as borane or lithium aluminum hydride to reduce both the ester and amide functions. The 3-(hydroxymethyl)-1-(arylmethyl)-2,3,4,5-tetrahydro-1H-[1,4]benzodiazepines 23 are oxidized with standard reagents known to convert a hydroxymethyl group to a carboxylic acid:reagents such as NaIO4 with catalyst RuO2 (e.g., see J. Org. Chem., 46:3936 (1981); Synlett, p. 143, (1996)). Coupling the acids (via the acid chlorides) to hydroxylamine then gives products 21. Certain intermediates as exemplified by formula 25 may be reduced with borane under mild conditions to give derivatives 25a in which the amide carbonyl is selectively reduced. These intermediates 25a are then converted to hydroxamic acid derivatives via hydrolysis of the ester to the acid and coupling the acid chloride with hydroxylamine.

 
wherein

 

 
R9 and R10 are: Cl, Br, F, OCH3, OEt , SCH3,

 

 
CF3, OCF3,

 
xe2x80x94NHCOCH3, or Me2Nxe2x80x94.

 
wherein R8=alkyl, arylalkyl, aryloxyalkyl, heterocyclicalkyl, or alkyloxyalkyloxyalkyl.
Other, preferred compounds of the present invention are those with basic moieties in the 1-(substituted carbonyl) group which may be prepared in the manner shown in Reaction Scheme 5. Reaction of the 2,3,4,5-tetrahydro-1H-[1,4]-benzodiazepines 14 (without a substituent at the 1-position) with carbonyl chloride derivatives in the manner depicted in Reaction Scheme 5, results in intermediates 25 which are then converted to acid 26 and hydroxamic acids 27. The intermediates 25 may also be synthesized by reaction of methyl 2-[(2-aminobenzyl)-(4-methoxybenzenesulfonyl)amino]-3-hydroxypropionates 6 with acid chlorides to give xe2x80x9cdehydroalaninexe2x80x9d derivatives 28. As previously described, mild bases such as NaHCO3 can be reacted with these derivatives to cause ring closure via a 1,4-addition to the double bond in intermediate 28 to provide the 7-membered 2,3,4,5-tetrahydro-1H-[1,4] diazepines 25.
As illustrated in Reaction Scheme 6, aryl-arylcarbonyl, heteroarylarylcarbonyl, aryl-heteroarylcarbonyl, heteroaryl-heteroarylcarbonyl derivatives 30 may be synthesized by standard palladium catalysed coupling of bromoaroyl or bromheteroaroyl derivatives 29 with appropriate arylstannanes, heteroarylstannanes, arylboronic acids, heteroarylboronic acids, aryl triflates, heteroaryl triflates and the like, under known conditions. For example, see Synthesis, 563-566 (1997); J. Org. Chem., 62:3405-3406, (1997); Tetrahedron Lett., 36:5247-5250, (1995); Heterocycles, 45:467, (1997); Tetrahedron Lett., 38:1118-1182, (1997); Heterocycles, 42:189-194, (1996); Tetrahedron Lett., 5005-5006, (1993); Synthesis, 843, (1987); Heterocycles, 2711-2716, (1987); and Tetrahedron Lett., 4407-4410, (1986).
By coupling with such palladium catalysts, aryl-aryl, heteroaryl-aryl, aryl-heteroaryl and heteroaryl-heteroaryl carboxylic ester derivatives can be prepared and these derivatives converted to carboxylic acid intermediates. The acids are then converted to acid chlorides which are reacted with esters of 2-[(2-aminobenzyl)-(4-substituted-benzenesulfonyl)amino]-3-hydroxypropionate as illustrated for conversion of derivatives 6 to intermediates 31.
The following references describe procedures for the synthesis of methyl 3-arylpyrrole-4-carboxylates as in J. Org. Chem., 62:2649-2651, (1997); methyl (2-methylphenyl) benzoates as in J. Org. Chem., 62:3405-3406, (1997); and methyl benzoates substituted with heterocyclic moieties such as furanyl, thienyl or pyridinyl groups as in Tetrahedron Lett., 27:4407-4410, (1986).

 
X is halogen, hydrogen, or (C1-C3)alkyl;
R and Rxe2x80x2 are as defined herein; and
R4 is as defined herein.

 
Y is H, F, Cl, CF3, CH3, or OCH3;
X is halogen, hydrogen, or (C1-C3)alkyl;
R and Rxe2x80x2 are as defined herein; and
R4 is as defined herein.
The intermediates 2,4,5,6-tetrahydro-1H-[1,4]benzodiazepines 39 and 38 may be prepared from glycine esters in the manner exemplified in Reaction Scheme 7. In this synthetic route, N-(4-substituted-benzenesulfonyl) derivatives of glycine ethyl ester, glycine t-butyl ester or glycine methyl ester 33 are alkylated with a substituted (R) or unsubstituted (Rxe2x95x90H) 2-nitrobenzyl bromide in N,N-dimethylformamide or 1-methyl-2-pyrrolidinone in the presence of potassium carbonate to give intermediates 34. Alternatively, the esters of N-(4-substituted-benzenesulfonyl) glycines, such as the methyl ester 33, are first reacted with sodium hydride in N,N-dimethylformamide or 1-methyl-2-pyrrolidinone and the resulting anion reacted with substituted or unsubstituted 2-nitrobenzylbromides to provide compounds 34. Reaction of derivates 34 with N,N-dimethyl(methylene)ammonium chloride or the iodide salts under standard reaction conditions (e.g., as set forth in Fieser and Fieser, 10:160-161; 8:194 affords the dimethylaminomethyl (Mannich type) compounds as intermediates for elimination to the xe2x80x9cdehydroalaninexe2x80x9d derivatives 37 or direct ring closure of 36 to 39 via an elimination-addition reaction. Ring closure of compounds 37 provides intermediates 38 for conversion to hydroxamic acids. Variations of the reactions conditions for conversion of 36 to 39 involve heating in the presence of Lewis acids, such as BF3, or heating an acid salt of 36 to effect the elimination-addition reaction.

 
The intermediate carboxylic acids for conversion to the tetrahydro[1,4]benzodiazepine-3-carboxylic acid, hydroxyamides may be synthesized via different routes as shown in Schemes 1-8. For the synthesis of some of the desired products of Formula 1, alternate routes are preferred as shown in Scheme 8. These routes may be preferred when the R4 group contains a triple bond or when it is preferred to introduce the R4 group toward the end of the synthetic sequence. Under these conditions, intermediate carboxylate esters of formula 41 or acids of formula 44 wherein the R4 substituent is an OH group are prepared. Intermediates with R4 an OH group any be prepared from derivatives wherein the OH group is protected by a group which can be selectively removed. Deriviatives 40 wherein R4 is an OCH3 moiety are suitable precurssors to the desired phenolic compounds 41 and 44 through cleavage of the oxygen methyl bond. As shown in Scheme 8 the anion of the phenolic OH group may be prepared in situ and then alkylated. Suitable bases are alkaline metal carbonates, hydrides, alkoxides and organic bases. Reaction with an alkylating moiety represented by the formula R15CH2X wherein X is a reactive leaving group such as a chloride,bromide, iodide, O-mesylate or an O-tosylate gives the derivatives 42 and 45.
The alkylation reaction may be carried out with carboxylate esters such as 41 or with the carboxylic acids represented by formula 44. Alternatively, the phenolic compounds 41 and 44 may be reacted under Mitsunobe Reaction conditions to afford the O-alkylated derivatives 42 and 45. Standard Mitsunobe Reaction conditions, which are described in the following literature references, may be used in the coupling reactions.
(a) J. Heterocyclic Chem. 34, 349 (1997);(b) Tetrahedron Lett 37, 6439 (1996);(c) J. Org Chem, 56, 7173 (1991);(d) Tetrahedron Lett 5709 (1989;(e) Synthesis 1-28 (1981).

 
The compounds of the present invention which have a basic moiety may be used in the form of salts derived from pharmaceutically or physiologically acceptable acids. These salts include, but are not limited to, salts with inorganic acids (such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid) or organic acids (such as acetic acid, oxalic acid, succinic acid, and maleic acid). Other salts of compounds with an acidic moiety include those with alkali metals or alkaline earth metals (such as sodium, potassium, calcium, and magnesium) or organic bases.
When the present compounds are utilized in pharmaceutical compositions, they may be combined with one or more pharmaceutically acceptable carriers, e.g., solvents, diluents and the like. Such compositions containing the present compounds may be administered orally, in the form of tablets, capsules, dispersible powders, granules, suspensions, syrups or elixirs; parentally, in the form of a sterile injectable solution or suspension; or topically, in the form of creams, lotions, ointments, etc. Such pharmaceutical compositions may contain from about 1 to about 100 mg of active ingredient in combination with the carrier.
The effective dosage of the present compounds utilized to treat a specific condition will vary depending upon the particular compound employed, the mode of administration and the type and severity of the condition being treated. However, in general, satisfactory results are obtained when the present compounds are administered at a dosage of about 0.001 to 1000 mg/kg of body weight.
As noted above, the compounds of the present invention may be administered orally, as well as by intravenous, intramuscular, subcutaneous or topical routes. Solid carriers useful for preparing tablets, capsules, etc., include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin. Liquid carriers useful for preparing compositions of the present compounds include sterile water, polyethylene, glycols, non-ionic surfactants, and edible oils such as corn, sesame, and peanut oils. Adjuvants conventionally used in the preparation of pharmaceutical compositions may also be included, such as flavoring agents, coloring agents, preservatives and antioxidants.
The compounds of the present invention were tested for biological activity according to the following procedures.
In Vitro Gelatinase Assay
The assay is based on the cleavage of the thiopeptide substrate ((Ac-Pro-Leu-Gly(2-mercapto-4-methyl-pentanoyl)-Leu-Gly-OEt), available from Bachem Bioscience) by the enzyme gelatinase, releasing the substrate product which reacts calorimetrically with DTNB ((5,5xe2x80x2-dithio-bis(2-nitro-benzoic acid)). This assay is disclosed in Weingarten et al., xe2x80x9cSpectrophotometric Assay for Vertebrate Collegenasexe2x80x9d, Anal. Biochem., 147:437-440, (1985). The enzyme activity is measured by the rate of the color increase.
The thiopeptide substrate was made up fresh as a 20 mM stock in 100% DMSO and the DTNB was dissolved in 100% DMSO as a 100 mM stock and stored in the dark at room temperature. The substrate and the DTNB were diluted together to 1 mM with substrate buffer (50 mM HEPES, pH 7.5, 5 mM CaCl2) before use. The stock of human neutrophil gelatinase B was diluted with assay buffer (50 mM HEPES, pH 7.5, 5 mM CaCl2, 0.02% Brij) to a final concentration of 0.15 nM.
The assay buffer, enzyme, DTNB/substrate (500 xcexcM final concentration) and vehicle or inhibitor were added to a 96 well plate (total reaction volume of 200 xcexcl) and the increase in color was monitored spectrophotometrically for 5 minutes at 405 nm on a plate reader.
The increase in OD405 was plotted and the slope of the line was calculated. The slope represents the reaction rate. The linearity of the reaction rate was confirmed (r2 greater than 0.85) and the mean (xxc2x1sem) of the control rate was calculated and compared for statistical significance (p less than 0.05) with drug-treated rates using Dunnett""s multiple comparison test. Dose-response relationships were generated using multiple doses of drug and IC50 values with 95% CI were estimated using linear regression (IPRED, HTB).
In Vitro Collagenase Assay
This assay was based on the cleavage of a peptide substrate ((Dnp-Pro-Cha-Gly-Cys(Me)-His-Ala-Lys(NMa)-NH2), available from Peptide International, Inc.) by collagenase releasing the fluorescent NMa group which was quantitated on the fluorometer as disclosed in Bickett et al., xe2x80x9cA High Throughput Fluorogenic Substrate for Interstitial Collagenase (MMP-1) and Gelatinase (MMP-9)xe2x80x9d, Anal. Biochem., 212:58-64, (1993). Dnp quenches the NMa fluorescence in the intact substrate.
The assay was run in HCBC assay buffer (50 mM HEPES, pH 7.0, 5 mM Ca+2, 0.02% Brij, 0.5% Cysteine), with human recombinant fibroblast collagenase (truncated, mw=18,828, from Wyeth-Ayerst Research, Radnor, Pa.). The substrate was dissolved in methanol and stored frozen in 1 mM aliquots. Collagenase was stored frozen in buffer in 25 xcexcM aliquots. In conducting the assay, the substrate was dissolved in HCBC buffer to a final concentration of 10 xcexcM and collagenase to a final concentration of 5 nM. The compounds being examined were dissolved in methanol, DMSO, or HCBC. The methanol and DMSO were diluted in HCBC to  less than 1.0%. The compounds were added to a 96 well plate containing enzyme and the reaction was started by the addition of substrate.
The reaction was read (excitation 340 nm, emission 444 nm) for 10 min. and the increase in fluorescence over time was plotted as a linear line. The slope of the line was calculated representing the reaction rate. The linearity of the reaction rate was confirmed (r2 greater than 0.85). The mean (xxc2x1sem) of the control rate was calculated and compared for statistical significance (p less than 0.05) with drug-treated rates using Dunnett""s multiple comparison test. Dose-response relationships were generated using multiple doses of drug and IC50 values with 95% CI were estimated using linear regression.
Procedure for Measuring TACE Inhibition
In a 96-well black microtiter plate, each well received a solution composed of 10 xcexcL TACE (available from Immunex) at a final concentration of 1 xcexcg/mL, 70 xcexcL Tris buffer, have a pH of 7.4 and containing 10% glycerol (final concentration 10 mM), and 10 xcexcL of test compound solution in DMSO (final concentration 1 xcexcM, DMSO concentration  less than 1%). The plates were incubated for 10 minutes at room temperature. The reaction was initiated by addition of a fluorescent peptidyl substrate (final concentration 100 xcexcM) to each well with shaking on a shaker for 5 sec.
The reaction was read (excitation 340 nm, emission 420 nm) for 10 min. and the increase in fluorescence over time was plotted as a linear line. The slope of the line was calculated and this represents the reaction rate. The linearity of the reaction rate was confirmed (r2 greater than 0.85). The mean (xxc2x1sem) of the control rate was calculated and compared for statistical significance (p less than 0.05) with drug-treated rates using Dunnett""s multiple comparison test. Dose-response relationships were generated using multiple doses of drug and IC50 values with 95% CI were estimated using linear regression.
Some results obtained following these standard experimental test procedures are present in Table 1.
 
Some intermediate compounds for the synthesis of derivatives of formula 1 are presented in Table 2 (Reference Examples 10,11,41,43-90,92-97,99,100,162,166,176,181,182,186,188,190)
 